Characterization of Senescence in Lemna gibba G3: A Determinate Growth System

Frond senescence in Lemna gibba G3 was characterized, and itscontrol by light, ABA and kinetin investigated. The plant exhibitsa determinate growth pattern with a frond producing a set numberof daughter fronds before undergoing senescence and death regardlessof whether or not it flowers. When a frond was cut in half,the distal half (half frond) which lacks any meristem underwentrapid senescence as compared with intact fronds. In both intactand half fronds, the onset of senescence was accelerated byABA and retarded by kinetin. Continuous white light acceleratedsenescence in both intact and half fronds over the dark controls.Under different photoperiodic light regime, the pace of daughterfrond production is accelerated in proportion to the lengthof light period. In half fronds, however, very short photoperiodiclight treatments (e.g. 1L: 23D or 3L: 21D) rather delayed senescenceover the dark controls. Two separate light control systems operatingin opposite directions in Lemana senescence appear to exist. ¹Present address: Department of Biology, Yonsei University,Seoul 120-749, Korea ²Present address: U.S. Department of Agriculture, Aero SpaceBuilding, Rm. 323, 901 D Street, S.W., Washington, D.C. 20251-2200, U.S.A. (Received July 13, 1989; Accepted May 8, 1990)     

Molecular genetic study of human arginase deficiency

We have explored the molecular pathology in 28 individuals homozygous or heterozygous for liver arginase deficiency (hyperargininemia) by a combination of Southern analysis, western blotting, DNA sequencing, and PCR. This cohort represents the majority of arginase-deficient individuals worldwide. Only 2 of 15 homozygous patients on whom red blood cells were available had antigenically cross-reacting material as ascertained by western blot analysis using anti-liver arginase antibody. Southern blots of patient genomic DNAs, cut with a variety of restriction enzymes and probed with a near-full-length (1,450-bp) human liver arginase cDNA clone, detected no gross gene deletions. Loss of a TaqI cleavage site was identified in three individuals: in a homozygous state in a Saudi Arabian patient at one site, at a different site in homozygosity in a German patient, and in heterozygosity in a patient from Australia. The changes in the latter two were localized to exon 8, through amplification of this region by PCR and electrophoretic analysis of the amplified fragment after treatment with TaqI; the precise base changes (Arg291X and Thr290Ser) were confirmed by sequencing. It is interesting that the latter nucleotide variant (Thr290Ser) was found to lie adjacent to the TaqI site rather than within it, though whether such a conservative amino acid substitution represents a true pathologic mutation remains to be determined. We conclude that arginase deficiency, though rare, is a heterogeneous disorder at the genotypic level, generally encompassing a variety of point mutations rather than substantial structural gene deletions.     

Characterization of "depolarization"-induced calcium release from sarcoplasmic reticulum in vitro with the use of membrane potential probe.

In the triad, the complex of transverse (T) tubule and sarcoplasmic reticulum (SR) Ca2+ release is induced from SR by mediation of the T-tubule. We report here evidence that this Ca2+ release is produced by depolarization of the T-tubule moiety. Thus, we found that the amount of [14C]SCN- taken up by T-tubules and triads (but not that by SR) increased upon incubation with (K, Na) gluconate, Mg ATP, indicating that the T-tubule was polarized making the lumenal side (equivalent to the extracellular side of an intact muscle fiber) more positive. Upon mixing with choline chloride, the procedure to induce Ca2+ release, [14C]SCN- uptake decreased, indicating that the T-tubule became depolarized. Activation of the T-tubule polarization by Na+ and prevention of it by digoxin [inhibitor of the (Na+, K+) pump], respectively, led to activation and inhibition of choline chloride-induced SR Ca2+ release.     

Kinetics of producing pro-urokinase in perfusion cultivations

Summary Better production of pro-urokinase from human cell line was observed with 5% serum containing medium than 10% or serum free medium on Cytodex II under perfusion chemostat operations, showing 0.8×10^–5 (IU/daycell) of maximum productivity at 0.020 (l/h) of dilution rate in 5% serum medium, which corresponds to 800 IU/mL at this dilution rate. Conversion of pro-urokinase was reduced in the serum-containing media.     

31例非特异性阴道炎病人阴道微生物群的研究

本文对31例非特异性阴道炎病人和31例正常人阴道微生物群进行定性、定量分析.结果表明,阴道炎病人的阴道乳杆菌的平均浓度明显低于正常人,而肠杆菌科、葡萄球菌、棒状杆菌、类杆菌属、支原体却明显增多,阴道炎病人未发现特异性病原体,菌群失调可能是其发病因素。乳杆菌为阴道正常优势菌,对改善阴道的微环境,防止条件致病菌引起的内源性感染具有重要的生理作用。     

Genetic Interactions between Reg1/Hex2 and Glc7, the Gene Encoding the Protein Phosphatase Type 1 Catalytic Subunit in Saccharomyces Cerevisiae

Mutations in GLC7, the gene encoding the type 1 protein phosphatase catalytic subunit, cause a variety of abberrant phenotypes in yeast, such as impaired glycogen synthesis and relief of glucose repression of the expression of some genes. Loss of function of the REG1/HEX2 gene, necessary for glucose repression of several genes, was found to suppress the glycogen-deficient phenotype of the glc7-1 allele. Deletion of REG1 in a wild-type background led to overaccumulation of glycogen as well as slow growth and an enlarged cell size. However, loss of REG1 did not suppress other phenotypes associated with GLC7 mutations, such as inability to sporulate or, in cells bearing the glc7(Y-170) allele, lack of growth at 14°. The effect of REG1 deletion on glycogen accumulation is not simply due to derepression of glucose-repressed genes, although it does require the presence of SNF1, which encodes a protein kinase essential for expression of glucose-repressed genes and for glycogen accumulation. We propose that REG1 has a role in controlling glycogen accumulation.     

小麦×玉米杂交后代的蛋白质及酯酶同工酶分析

以８ 个普通小麦（Ｔｒｉｔｉｃｕｍ ａｅｓｔｉｖｕｍ Ｌ．）品种为母本,２ 个栽培玉米（Ｚｅａ ｍ ａｙｓ Ｌ．）品种为父本杂交所获得的Ｆ２ 代在形态上出现了明显变异。对其籽粒进行蛋白质电泳分析,得到了如下主要结果：杂交后代的蛋白质谱带较母本有了很大的变异,主要集中在高分子量麦谷蛋白（ＨＭＷ－Ｇｌｕ）区域。杂交后代的蛋白质谱带由５ 种类型构成：１．母本型,占全部测试籽粒的２２．６％ ;２．附加型,占１４．３％ ;３．互补型,占１５．５％ ;４．杂种型,占３０．９％ ;５．缺失型,占１６．７％ 。对“矮杆早”ד紫粘”的Ｆ２ 代籽粒进行酯酶同工酶电泳分析发现,变异主要发生在ＥＳＴ－１ 区。由此看来,小麦×玉米的杂合子中玉米染色体在被排除前后,可以诱发小麦染色体组发生遗传变异     

Production of a novel copolyester of 3-hydroxybutyric acid and medium-chain-length 3-hydroxyalkanoic acids by Pseudomonas sp. 61-3 from sugars

 Pseudomonas sp. 61-3 (isolated from soil) produced a polyester consisting of 3-hydroxybutyric acid (3HB) and of medium-chain-length 3-hydroxyalkanoic acids (3HA) of C₆, C₈, C₁₀ and C₁₂, when sugars of glucose, fructose and mannose were fed as the sole carbon source. The polyester produced was a blend of homopolymer and copolymer, which could be fractionated with boiling acetone. The acetone-insoluble fraction of the polyester was a homopolymer of 3-hydroxybutyrate units [poly (3HB)], while the acetone-soluble fraction was a copolymer [poly(3HB-co-3HA)] containing both short- and medium-chain-length 3-hydroxyalkanoate units ranging from C₄ to C₁₂:44 mol% 3-hydroxybutyrate, 5 mol% 3-hydroxyhexanoate, 21 mol% 3-hydroxyoctanoate, 25 mol% 3-hydroxydecanoate, 2 mol% 3-hydroxydodecanoate and 3 mol% 3-hydroxy-5-cis-dodecenoate. The copolyester was shown to be a random copolymer of 3-hydroxybutyrate and medium-chain-length 3-hydroxyalkanoate units by analysis of the ¹³C-NMR spectrum. The poly(3HB) homopolymer and poly (3HB-co-3HA) copolymer were produced simultaneously within cells from glucose in the absence of any nitrogen source, which suggests that Pseudomonas sp. 61-3 has two types of polyhydroxy-alkanoate syntheses with different substrate specificities. Received: 9 June 1995/Received last revision: 30 October 1995/Accepted: 6 November 1995     

Charactererization of novel non-toxic Bacillus thuringiensis isoloates from Korea

J.Y. ROH, H.W. PARK, B.R. JIN, H.S. KIM, Y.M. YU AND S.K. KANG. 1996. Four Bacillus thuringiensis isolates from soil samples produced parasporal inclusions which were non-toxic to insects. The isolates were named B. thuringiensis NTB-1, NTB-2, NTB-3 and NTB-4. The parasporal inclusions were shown to be ovoid by phase contrast and scanning electron microscopy. The serotypes of the four isolates were determined by agglutination using 33 antisera; NTB-1 and NTB-4 seemed to be subsp. isruelensis ,and NTB-2 seemed to be subsp. pondzcheriensis . NTB-3 did not react with the 33 antisera. However, comparison of parasporal protein and plasmid DNA patterns of the four isolates with those of 15 known non-toxic B. thuringiensis strains demonstrated that the four isolates are novel.     

Ras induces anchorage-independent growth by subverting multiple adhesion-regulated cell cycle events.

Anchorage-independent growth is a hallmark of transformed cells, but little is known of the molecular mechanisms that underlie this phenomenon. We describe here studies of cell cycle control of anchorage-independent growth induced by the ras oncogene, with the use of a somatic cell mutant fibroblast line (ER-1-2) that is specifically defective in oncogene-mediated, anchorage-independent growth. Control, nontransformed PKC3-F4 cells and ER-1-2 cells cannot proliferate in semisolid medium. Three important cell cycle events are dependent on adhesion of these cells to a substratum: phosphorylation of the retinoblastoma protein, pRB; cyclin E-dependent kinase activity; and cyclin A expression. PKC3-F4 cells that express ras (PKC3-F4/ras cells) proliferate in nonadherent cultures, and each of these three events occurs in the absence of adhesion in PKC3-F4/ras cells. Thus, ras can override the adhesion requirement of cellular functions that are necessary for cell cycle progression. ER-1-2 cells that express ras (ER-1-2/ras cells) possess hyperphosphorylated forms of pRB and cyclin E-dependent kinase activity in the absence of adhesion but remain adhesion dependent for expression of cyclin A. The adhesion dependence of pRB phosphorylation and cyclin E-dependent kinase activity is therefore dissociable from the adhesion dependence of cyclin A expression. Furthermore, ectopic expression of cyclin A is sufficient to rescue anchorage-independent growth of ER-1-2/ras cells but does not induce anchorage-independent growth of PKC3-F4 or ER-1-2 cells. However, like pRB phosphorylation and cyclin E-dependent kinase activity, the kinase activity associated with ectopically expressed cyclin A is dependent on cell adhesion, and this dependence is overcome by ras. Thus, the induction of anchorage-independent growth by ras may involve multiple signals that lead to both expression of cyclin A and activation of G1 cyclin-dependent kinase activities in the absence of cell adhesion.